Mitochondrial Dysfunction in Skeletal Muscle of Rotenone-Induced Rat Model of Parkinson's Disease: SC-Nanophytosomes as Therapeutic Approach.
The development of new therapeutic options for Parkinson's disease (PD) requires formulations able to mitigate both brain degeneration and motor dysfunctions. SC-Nanophytosomes, an oral mitochondria-targeted formulation developed with Codium tomentosum membrane polar lipids and elderberry anthocyanin-enriched extract, promote significant brain benefits on a rotenone-induced rat model of PD. In the present work, the effects of SC-Nanophytosome treatment on the skeletal muscle tissues are disclosed. It is unveiled that the rotenone-induced PD rat model exhibits motor disabilities and skeletal muscle tissues with deficient activity of mitochondrial complexes I and II along with small changes in antioxidant enzyme activity and skeletal muscle lipidome. SC-Nanophytosome treatment mitigates the impairment of complexes I and II activity, improving the mitochondrial respiratory chain performance at levels that surpass the control. Therefore, SC-Nanophytosome competence to overcome the PD-related motor disabilities should be also associated with its positive outcomes on skeletal muscle mitochondria. Providing a cellular environment with more reduced redox potential, SC-Nanophytosome treatment improves the skeletal muscle tissue's ability to deal with oxidative stress stimuli. The PD-related small changes on skeletal muscle lipidome were also counteracted by SC-Nanophytosome treatment. Thus, the present results reinforces the concept of SC-Nanophytosomes as a mitochondria-targeted therapy to address the neurodegeneration challenge.


Sensitivity of 14-3-3 protein test varies in subtypes of sporadic Creutzfeldt-Jakob disease.
The increase of the 14-3-3 protein in CSF is used as a diagnostic test in Creutzfeldt-Jakob disease (CJD), but the sensitivity and specificity of the 14-3-3 test are disputed. One reason for the dispute may be the recently established heterogeneity of sporadic CJD. The relationship between CSF 14-3-3 protein and sporadic CJD subtypes, distinguished by electrophoretic mobility of proteinase K-resistant prion protein (PrP(Sc)) and genotype at codon 129 of the prion protein gene, has not been elucidated. The authors examined the 14-3-3 protein test in 90 patients with sporadic CJD. PrP(Sc) type (type 1 or type 2) and the genotype at polymorphic codon 129 were determined in each patient. Mutations were excluded by prion gene sequencing. The authors' findings indicate that the sensitivity of the 14-3-3 test is higher in patients with molecular features of the classic sporadic CJD than in patients with the nonclassic CJD subtypes. The difference appears to be related to the PrP(Sc) type and not to the codon 129 genotype. Disease duration before 14-3-3 testing might also have an influence because it was shorter in classic sporadic CJD. The Creutzfeldt-Jakob disease clinical subtype should be considered when interpreting results of the 14-3-3 test.


Tackling HIV in India: evidence-based priority setting and programming.
In the wake of a downward revision of the number of HIV-infected people, India is launching an ambitious US$2.5 billion, five-year HIV plan. Responding to new data on HIV prevalence and risk behavior, India has earmarked almost 70 percent of the budget for prevention; one-third focuses on prevention activities for those at highest risk of HIV, and the remainder addresses HIV testing expansion and services for pregnant women. About 20 percent of the total budget is for care and treatment. Although the size and scope of the proposed HIV response pose challenges, the world has much to learn from India's data-informed approach to policy and priority setting.


Cold exposure or chronic noradrenaline treatment induces an increase in the calmodulin-like immunoreactivity of brown adipose tissue of rats.
Cell proliferation is often associated with an increase in calmodulin, the ubiquitous intracellular calcium receptor of non-muscle cells. A long lasting increase in the proliferative activity of brown adipose tissue is induced by cold exposure in the rat. The present work showed that this phenomenon is also associated with a rapid and long lasting increase in the calmodulin content of this tissue. It was equally shown that this increase can be reproduced by noradrenaline administration.


Properties of zein films coated with drying oils.
Zein films prepared by resin were coated with either flax oil or tung oil and cured by UV- or gamma-radiation. Coated zein films were then evaluated for tensile and water barrier properties. Film microstructure was examined by transmitted light microscope. Tensile strength, elongation, and toughness of oil-coated samples increased substantially with respect to uncoated films. Flax oil coated samples showed an increase in elongation of 300%. It was suggested that oil coatings fill in pinholes and cracks existing in zein films, affecting their mechanical properties. Water vapor permeability also decreased statistically for coated film (except for tung oil coated-UV treated films), suggesting water vapor transfer was controlled by film hydrophobicity and microstructure. Moreover, the liquid water transmission rate of coated films was at least 10 times slower than for control films. Examination of film microstructure revealed that flax oil coatings had uniform coverage and smooth finish, which explained their high elongation, low water vapor permeability. Tung oil coatings cured under UV light showed patterns of intertwined dark and light regions, which may be caused by cross-linking and drying at different times. The oriented structures were found when tung oil coatings were cured by gamma-radiation.


Higher professional education for general medical practitioners: key informant interviews and focus group findings.
If higher professional education (HPE) for general practitioners (GPs) is to be implemented, then key stakeholders will need to be supportive. To investigate stakeholders' beliefs about the concept of HPE, its funding, and relationships to education and care. Interviews were conducted using a topic guide with a health authority (HA) representative, the Local Medical Committee Chair, the Medical Audit Advisory Group Chair, a GP tutor from each of the six health authorities in the old South West region, and a senior member of the three academic GP departments and the two Royal College of General Practitioners faculties in the region. Focus groups were held with GP registrars on both vocational training schemes (VTSs) and on the one HPE course in the region. These were recorded, transcribed verbatim, and analysed for emergent themes that were triangulated with the ideas expressed in the focus groups; the same topic guide was used for both. Of 29 key informants, 24 were interviewed. Six focus groups were held (the one HPE group and five out of the nine VTSs), after which no new ideas emerged. There is a transition period, after becoming a new principal (NP) and before becoming a fully competent independent GP, during which NPs need support. Benefits would include receiving peer support to reduce stress during the transition, enhanced non-clinical competencies, becoming a better skilled GP, avoiding the negative personal impact of a career as a GP, and helping recruitment. To improve patient care there must be a link between education and service provision. Funding is the major consideration in setting HPE; mixed funding is best coming from top-sliced General Medical Services (GMS), the HA, and regional educational funds. Barriers might include NPs' practice workload, their enthusiasm, and their partners' attitudes. The other key is a local enthusiast to initiate a course and coordinate the 'players'. The curriculum would be principally non-clinical and should be agreed by learners and the course tutor together, taking advice from various interested parties. There is a need for HPE for new NPs. It will require funding external to individual practices or NPs and a local enthusiast. Top-slicing of GMS funds is one source of funding, with additional funds from regional education and HAs. HPE must be related to service provision, to NP needs, and to vocational training.


Porphyrin-based sensitizers in the detection and treatment of cancer: recent progress.
It has been known for some time that porphyrins and related compounds have the ability to selectively accumulate in tumor tissues, and to persist there for long periods of time. This property, along with the well-described photophysical and photosensitizing properties of porphyrin-type molecules, has led to their potential use as adjuvants and sensitizers in a variety of medical applications, such as in photodynamic therapy (PDT), boron neutron capture therapy (BNCT), radiation therapy (RT) and in magnetic resonance imaging (MRI). Both PDT and BNCT are binary cancer therapies that involve activation of tissue-localized sensitizers with either light (in PDT) or low-energy neutrons (in BNCT). In both of these therapeutic methodologies, local tumor control with minimal side effects relative to other forms of cancer treatment (surgery, radiotherapy, chemotherapy) can be achieved. Porphyrins constitute a major class of pharmacological agents currently under investigation. Photofrin, a porphyrin derivative, has been approved in the USA as a PDT drug by the U.S. Food and Drug Administration (FDA), and also in Japan, Canada and in eleven European countries. Recently, the FDA approved Visudyne, another porphyrin derivative for the PDT treatment of the 'wet-form' of age-related macular degeneration. In addition to cancer treatment porphyrins are also under investigation for application in the treatment of a variety of other diseases.


Characterization of calcium transients during early embryogenesis in ascidians Ciona robusta (Ciona intestinalis type A) and Ciona savignyi.
The calcium ion (Ca2+) is an important second messenger, and a rapid increase in Ca2+ level (Ca2+ transient) is involved in various aspects of embryogenesis. Although Ca2+ transients play an important role in early developmental stages, little is known about their dynamics throughout embryogenesis. Here, Ca2+ transients were characterized by visualizing Ca2+ dynamics in developing chordate embryos using a fluorescent protein-based Ca2+ indicator, GCaMP6s in combination with finely tuned microscopy. Ca2+ transients were detected in precursors of muscle cells in the late gastrula stage. In the neurula stage, repetitive Ca2+ transients were observed in left and right neurogenic cells, including visceral ganglion (VG) precursors, and the duration of Ca2+ transients was 39±4s. In the early tailbud stage, Ca2+ transients were observed in differentiating precursors of nerve cord neurons. A small population of VG precursors showed rhythmical Ca2+ transients with a duration of 22±4s, suggesting a central pattern generator (CPG) origin. At the mid tailbud stage, Ca2+transients were observed in a wide area of epidermal cells and named CTECs. The number and frequency of CTECs increased drastically in late tailbud stages, and the timing of the increase coincided with that of the relaxation of the tail bending. The experiment using Ca2+ chelator showed that the CTECs were largely depending on the extracellular Ca2+. The waveform analysis of Ca2+ transients revealed different features according to duration and frequency. The comprehensive characterization of Ca2+ transients during early ascidian embryogenesis will help our understanding of the role of Ca2+ signaling in chordate embryogenesis.


Diversity and dynamics of fungi during spontaneous fermentations and association with unique aroma profiles in wine.
Microbial ecology is an integral part of an agricultural ecosystem and influences the quality of agricultural commodities. Microbial activity influences grapevine health and crop production, conversion of sugar to ethanol during fermentation, thus forming wine aroma and flavour. There are regionally differentiated microbial patterns in grapevines and must but how microbial patterns contribute to wine regional distinctiveness (terroir) at small scale (<100 km) is not well defined. Here we characterise fungal communities, yeast populations, and Saccharomyces cerevisiae populations during spontaneous fermentation using metagenomics and population genetics to investigate microbial distribution and fungal contributions to the resultant wine. We found differentiation of fungi, yeasts, and S. cerevisiae between geographic origins (estate/vineyard), with influences from the grape variety. Growth and dominance of S. cerevisiae during fermentation reshaped the fungal community and showed geographic structure at the strain level. Associations between fungal microbiota diversity and wine chemicals suggest that S. cerevisiae plays a primary role in determining wine aroma profiles at a sub-regional scale. The geographic distribution at scales of less than 12 km supports that differential microbial communities, including the dominant fermentative yeast S. cerevisiae can be distinct in a local setting. These findings provide further evidence for microbial contributions to wine terroir, and perspectives for sustainable agricultural practices to maintain microbial diversity and optimise fermentation function to craft beverage quality.


Development of an instrument evaluating the impact of surgeon-patient relationship in patients on sick leave.
Background: To date, no specific instruments exist to measure the quality of the patient-surgeon relationship despite its potential to influence clinical and economic outcomes in patients undergoing surgery for musculoskeletal disorders (MSDs). Objective: The objective was to develop and validate an instrument to assess the quality of the patient-surgeon relationship, taking into account the return to work after functional restoration surgery. Methods: The instrument development was based on literature review, cognitive interviews and expert examinations. The instrument's psychometric properties were explored in a sample of 50 French patients on sick leave with musculoskeletal disorders or hand injuries. Face validity, internal consistency and test-retest reliability were evaluated. The dimensionality of the instrument was studied using an exploratory principal component analysis. Results: The 11-item instrument showed good psychometric properties. The cognitive interviews allowed enhancing the validity of the instrument content by capturing patients' point of view. The exploratory principal component analysis demonstrated the uni-dimensionality of the instrument with the first factor accounting for 83% of the total explained variance. Conclusion:This study has developed the first instrument capable of the specific assessment of the impact of the surgeon-patient relationship on recovery, in patients with hand traumas and MSDs.


The Development and Implementation of a School-Based Dental Sealant Program for Hawaii Public Schools.
School-based dental sealant programs are noted to increase sealant uptake among children, especially those from low-income families and those living in rural and underserved areas. However, nationally school-based dental sealant programs are substantially underutilized. This paper describes a partnership approach to a school-based dental sealant program for Title 1 public schools in Hawaii. The Hawaii Keiki: Healthy and Ready to Learn program partnered with the Hawaii Dental Service to develop and implement an interprofessional, school-based sealant program (SBSP). This article describes the development of key factors, such as partnerships, an interprofessional team, team roles, policies, and marketing tools to optimize program success. The SBSP has completed over 3 academic years of implementation in Hawaii public schools. In its initial year (school year [SY] 2019-2020), which presented challenges due to the COVID-19 pandemic, it served 6 schools (106 students screened) on 1 island. It expanded to 14 schools on 1 island (193 students screened) in its second year (SY 2020-2021), 28 schools on 3 islands (653 students screened) in its third year (SY 2021-2022), and during the Fall of the fourth year, the program reached 50 schools on 3 islands (1821 students screened) and is projected to reach over 70 schools by the end of SY 2022-2023. Taking advantage of strategic partnerships and key interprofessional team members, a school-based dental sealant program can help to improve the oral health of children in public schools by helping to decrease barriers to care.


HCN subunit-specific and cAMP-modulated effects of anesthetics on neuronal pacemaker currents.
General anesthetics have been a mainstay of surgical practice for more than 150 years, but the mechanisms by which they mediate their important clinical actions remain unclear. Ion channels represent important anesthetic targets, and, although GABA(A) receptors have emerged as major contributors to sedative, immobilizing, and hypnotic effects of intravenous anesthetics, a role for those receptors is less certain in the case of inhalational anesthetics. The neuronal hyperpolarization-activated pacemaker current (Ih) is essential for oscillatory and integrative properties in numerous cell types. Here, we show that clinically relevant concentrations of inhalational anesthetics modulate neuronal Ih and the corresponding HCN channels in a subunit-specific and cAMP-dependent manner. Anesthetic inhibition of Ih involves a hyperpolarizing shift in voltage dependence of activation and a decrease in maximal current amplitude; these effects can be ascribed to HCN1 and HCN2 subunits, respectively, and both actions are recapitulated in heteromeric HCN1-HCN2 channels. Mutagenesis and simulations suggest that apparently distinct actions of anesthetics on V(1/2) and amplitude represent different manifestations of a single underlying mechanism (i.e., stabilization of channel closed state), with the predominant action determined by basal inhibition imposed by individual subunit C-terminal domains and relieved by cAMP. These data reveal a molecular basis for multiple actions of anesthetics on neuronal HCN channels, highlight the importance of proximal C terminus in modulation of HCN channel gating by diverse agents, and advance neuronal pacemaker channels as potentially relevant targets for clinical actions of inhaled anesthetics.


Endoscopic Endonasal Surgery for Dermoid Cysts Arising From the Middle Cranial Fossa Floor: A Rare Case Series.
Dermoid cysts (DCs) are congenital, slowly growing, and may cause nervous system symptoms. Related literature is limited and mainly includes case reports. We report a case series of DCs originating from the middle cranial fossa floor (MCFF) and investigate their demographic information, clinical characteristics, imaging findings, surgical procedures, and prognostic outcomes. We reviewed the patients with DCs arising from the MCFF undergoing endoscopic endonasal surgery (EES) in our center between 2012 and 2022. A total of 5 patients with DCs were enrolled (2 males and 3 females), with a mean age of 46.2 years at the onset. All DCs originated from the MCFF with 1 case involving the middle cranial fossa bone and another 1 case affecting the dura mater. One (20.0%) patient had neurological involvement. After admission, all patients received EES with a total resection rate of 100.0% (5 of 5). After a median follow-up of 73.2 months, all patients achieved complete clinical and radiological improvements. No surgical-related complications or relapses were observed during the long-term follow-up. Endoscopic endonasal surgery is considered a safe and effective approach for the treatment of DCs in the MCFF. A larger sample size and longer follow-up time are needed.


Identity of a Rust on Ephedra.
The aecial stage of a rust on Ephedra has been called a Peridermium. Clues as to its telial stage have been lacking. It is here described as resembling more a form of Roestelia, and the reasons for its possible relation to Gymnosporangium multiporum Kern are presented: the two sets of hosts have similar distributions: the two rust stages have the same geographic range; they have been found in close proximity in the field.


Anatomical evidence for the involvement of the macaque ventrolateral prefrontal area 12r in controlling goal-directed actions.
The macaque ventrolateral prefrontal (VLPF) area 12r is thought to be involved in higher-order nonspatial information processing. We found that this area is connectionally heterogeneous, and the intermediate part is fully integrated in a cortical network involved in selecting and controlling object-oriented hand and mouth actions. Specifically, intermediate area 12r displayed dense connections with the caudal half of area 46v and orbitofrontal areas and relatively strong extraprefrontal connections involving the following: (1) the hand- and mouth-related ventral premotor area F5 and the anterior intraparietal (AIP) area, jointly involved in visuomotor transformations for grasping; (2) the SII sector that is connected to AIP and F5; (3) a sector of the inferotemporal area TEa/m, primarily corresponding to the sector densely connected to AIP; and (4) the insular and opercular frontal sectors, which are connected to AIP and F5. This connectivity pattern differed markedly from those of the caudal and rostral parts of area 12r. Caudal area 12r displayed dense connections with the caudal part of the VLPF, including oculomotor areas 8/FEF and 45B, relatively weak orbitofrontal connections and extraprefrontal connections limited to the inferotemporal cortex. Rostral area 12r displayed connections mostly with rostral prefrontal and orbitofrontal areas and relatively weaker connections with the fundus and the upper bank of the superior temporal sulcus. The present data suggest that the intermediate part of area 12r is involved in nonspatial information processing related to object properties and identity, for selecting and controlling goal-directed hand and mouth actions.


[Surgical key points. Trabeculotomy].
Diagnosis and treatment of congenital glaucoma is an emergency. The objective of trabeculotomy is to eliminate the trabecular obstruction by creating a new drainage channel from Schlemm's canal. Performed on distended and fragile eyes, this surgical procedure requires great caution.


Mesenchymal stem cells protect breast cancer cells through regulatory T cells: role of mesenchymal stem cell-derived TGF-beta.
Mesenchymal stem cells (MSCs) have been shown to support breast cancer growth. Because MSCs also increase the frequency of regulatory T cells (T(regs)), this study tested the hypothesis that human MSCs, via Tregs, protect breast cancer cells (BCCs) from immune clearance MSCs suppressed the proliferation of PBMCs when the latter were exposed to gamma-irradiated BCCs. Similarly, MSCs showed significant inhibition of PBMC migration toward BCCs and a corresponding decrease in CXCL12. MSCs also inhibited NK cell and CTL functions, which correlated with reduced numbers of CD8(+) and CD56(+) cells compared with parallel cultures without MSCs. The reduced NK and CTL activities correlated with a decrease in intracellular and secreted granzyme B. To explain these immunosuppressive findings, we compared T(reg) levels after coculture with MSCs and found an approximately 2-fold increase in T(regs), with associated decreases in antitumor Th1 cytokines and increases in Th2 cytokines. MSC-derived TGF-beta1 was largely responsible for the increase in T(regs) based on knockdown studies. In the presence of T(reg) depletion, PBMC proliferation and effector functions were partially restored. Together, these studies show an MSC-mediated increase in T(regs) in cocultures of PBMCs and BCCs. The results could be explained, in part, by the increase in Th2-type cytokines and MSC-generated TGF-beta1. These findings demonstrate immune protection by MSCs to BCCs. The reduction in immune cell proliferation and recruitment mediated by MSCs has implications for treatment of breast cancer with chemotherapy.


Albumin conjugation promotes arsenic trioxide transport through alkaline phosphatase-associated transcytosis in MUC4 wildtype pancreatic cancer cells.
Pancreatic cancer (PC) has a poor prognosis due to chemotherapy resistance and unfavorable drug transportation. Albumin conjugates are commonly used as drug carriers to overcome these obstacles. However, membrane-bound glycoprotein mucin 4 (MUC4) has emerged as a promising biomarker among the genetic mutations affecting albumin conjugates therapeutic window. Human serum albumin-conjugated arsenic trioxide (HSA-ATO) has shown potential in treating solid tumors but is limited in PC therapy due to unclear targets and mechanisms. This study investigated the transport mechanisms and therapeutic efficacy of HSA-ATO in PC cells with different MUC4 mutation statuses. Results revealed improved penetration of ATO into PC tumors through conjugated with HSA. However, MUC4 mutation significantly affected treatment sensitivity and HSA-ATO uptake both in vitro and in vivo. Mutant MUC4 cells exhibited over ten times higher IC50 for HSA-ATO and approximately half the uptake compared to wildtype cells. Further research demonstrated that ALPL activation by HSA-ATO enhanced transcytosis in wildtype MUC4 PC cells but not in mutant MUC4 cells, leading to impaired uptake and weaker antitumor effects. Reprogramming the transport process holds potential for enhancing albumin conjugate efficacy in PC patients with different MUC4 mutation statuses, paving the way for stratified treatment using these delivery vehicles.


Gemcitabine plus Paclitaxel versus Paclitaxel monotherapy in patients with metastatic breast cancer and prior anthracycline treatment.
The objective of this phase III global study was to compare the efficacy of gemcitabine plus paclitaxel (GT) versus paclitaxel in patients with advanced breast cancer. It was designed as a pivotal study for the approval of G for a breast cancer treatment indication. Patients who relapsed after adjuvant anthracyclines were randomly assigned to gemcitabine,1,250 mg/m(2) days 1 and 8 plus paclitaxel, 175 mg/m(2) on day 1; or, to paclitaxel at same dose on day 1 (both arms administered every 21 days, unblinded). The primary end point was overall survival (OS) and secondary end points were time to progression (TTP), response rate (RR), progression-free survival, response duration, and toxicity. This final OS analysis was planned at 380 deaths. A total of 266 patients were randomly assigned to GT and 263 to paclitaxel. Median survival on GT was 18.6 months versus 15.8 months on paclitaxel (log-rank P = . 0489), with an adjusted Cox hazard ratio of 0.78 (95% CI, 0.64 to 0.96; P = .0187). The TTP was longer (6.14 v 3.98 months; log-rank P = .0002) and the RR was better (41.4% v 26.2%; P = .0002) on GT. There was more grade 3 to 4 neutropenia on GT and grade 2 to 4 fatigue and neuropathy were slightly more prevalent on GT. This phase III study documents a role for gemcitabine in advanced breast cancer after anthracycline-based adjuvant therapy. The results establish GT as a reasonable choice for women who require cytoreduction with manageable toxicities and validate ongoing testing of GT in the adjuvant setting.


Epilepsy and vaccinations: Italian guidelines.
Reports of childhood epilepsies in temporal association with vaccination have had a great impact on the acceptance of vaccination programs by health care providers, but little is known about this possible temporal association and about the types of seizures following vaccinations. For these reasons the Italian League Against Epilepsy (LICE), in collaboration with other Italian scientific societies, has decided to generate Guidelines on Vaccinations and Epilepsy. The aim of Guidelines on Vaccinations and Epilepsy is to present recent unequivocal evidence from published reports on the possible relationship between vaccines and epilepsy in order to provide information about contraindications and risks of vaccinations in patients with epilepsy. The following main issues have been addressed: (1) whether contraindications to vaccinations exist in patients with febrile convulsions, epilepsy, and/or epileptic encephalopathies; and (2) whether any vaccinations can cause febrile seizures, epilepsy, and/or epileptic encephalopathies. Diphtheria-tetanus-pertussis (DTP) vaccination and measles, mumps, and rubella vaccination (MMR) increase significantly the risk of febrile seizures. Recent observations and data about the relationships between vaccination and epileptic encephalopathy show that some cases of apparent vaccine-induced encephalopathy could in fact be caused by an inherent genetic defect with no causal relationship with vaccination.


High-Risk Breast Lesions: A Machine Learning Model to Predict Pathologic Upgrade and Reduce Unnecessary Surgical Excision.
Purpose To develop a machine learning model that allows high-risk breast lesions (HRLs) diagnosed with image-guided needle biopsy that require surgical excision to be distinguished from HRLs that are at low risk for upgrade to cancer at surgery and thus could be surveilled. Materials and Methods Consecutive patients with biopsy-proven HRLs who underwent surgery or at least 2 years of imaging follow-up from June 2006 to April 2015 were identified. A random forest machine learning model was developed to identify HRLs at low risk for upgrade to cancer. Traditional features such as age and HRL histologic results were used in the model, as were text features from the biopsy pathologic report. Results One thousand six HRLs were identified, with a cancer upgrade rate of 11.4% (115 of 1006). A machine learning random forest model was developed with 671 HRLs and tested with an independent set of 335 HRLs. Among the most important traditional features were age and HRL histologic results (eg, atypical ductal hyperplasia). An important text feature from the pathologic reports was "severely atypical." Instead of surgical excision of all HRLs, if those categorized with the model to be at low risk for upgrade were surveilled and the remainder were excised, then 97.4% (37 of 38) of malignancies would have been diagnosed at surgery, and 30.6% (91 of 297) of surgeries of benign lesions could have been avoided. Conclusion This study provides proof of concept that a machine learning model can be applied to predict the risk of upgrade of HRLs to cancer. Use of this model could decrease unnecessary surgery by nearly one-third and could help guide clinical decision making with regard to surveillance versus surgical excision of HRLs. © RSNA, 2017.


Genomic sequence of human glyoxalase-I: analysis of promoter activity and its regulation.
Glyoxalase-I is a glutathione-binding protein involved in the detoxification of methylglyoxal, a by-product of glycolysis. Aberrations in the expression of human glyoxalase in cancer and diabetes have been reported. To gain a better understanding of the glyoxalase-I regulation under normal physiological conditions and in disease processes, we have cloned 12kb of genomic sequence, comprising five exons, separated by four introns. A fragment comprising 982bp of 5' flanking region was used in the pSEAP reporter system to identify the minimal promoter and to locate any cis-acting functional elements. This region contained a minimal promoter between -20 and -160bp. Cells transfected with a construct containing the 5' flanking sequence exhibited a 45-fold higher activity over vector transfected cells. A twofold reproducible increase in reporter activity was seen with insulin and ZnCl(2) treatments, indicating a functionally operative insulin response element (IRE) and metal response element (MRE). Knowledge regarding the regulation of glyoxalase-I may provide insights into the importance of this enzyme in human diseases.


[Acute myeloblastosis in von Jaksch-Luzet syndrome].
A 7 month-old infant of Italian origin, who was given a vitamin-D deficient diet developed a Von Jaksch-Luzet syndrome with more than 20% myeloblasts in the bone marrow. Plasma levels of vitamin D metabolites wer low and parathormone levels were high. A low dose of vitamin D3 (2,000 IU cholecalciferol/day) resulted in the recovery of a normal phosphocalcic balance and in the disappearance of myeloblastosis.


Identification of a new common provirus integration site in gross passage A murine leukemia virus-induced mouse thymoma DNA.
The Gross passage A murine leukemia virus (MuLV) induced T-cell leukemia of clonal (or oligoclonal) origin in inoculated mice. To study the role of the integrated proviruses in these tumor cells, we cloned several newly integrated proviruses (with their flanking cellular sequences) from a single tumor in procaryotic vectors. With each of the five clones obtained, a probe was prepared from the cellular sequences flanking the provirus. With one such probe (SS8), we screened several Gross passage A MuLV-induced SIM.S mouse tumor DNAs and found that, in 11 of 40 tumors, a provirus was integrated into a common region designated Gin-1. A 26-kilobase-pair sequence of Gin-1 was cloned from two lambda libraries, and a restriction map was derived. All proviruses were integrated as a cluster in the same orientation within a 5-kilobase-pair region of Gin-1, and most of them had a recombinant structure of the mink cell focus-forming virus type. The frequency of Gin-1 occupancy by provirus was much lower in thymoma induced by other strains of MuLV in other mouse strains. Using somatic-cell hybrid DNAs, we mapped Gin-1 on mouse chromosome 19. Gin-1 was not homologous to 16 known oncogenes and was distinct from the other common regions for provirus integration previously described. Therefore, Gin-1 appears to represent a new common provirus integration region. The integration of a provirus within Gin-1 might be an important event leading to T-cell transformation, and the Gin-1 region might harbor sequences which are involved in tumor development.


Psychological characteristics and associations between living kidney transplantation recipients and biologically related or unrelated donors.
Although recipients and donors in living kidney transplantation experience psychological distress-including depression and anxiety-during the pre-operative period, very few studies have evaluated the related psychological reactions. This study aimed to determine the characteristics and correlations of the mood states and personality of recipients and donors (genetically related and unrelated) of living kidney transplantations. A total of 66 pairs of living donors and recipients were enrolled from April 2008 to June 2019 in this study, of whom 53 eligible pairs of living donors and recipients were included in the retrospective analysis of their psychological assessments in the pre-transplantation states. While participants' personality patterns were assessed using the Minnesota Multiphasic Personality Inventory-2 (MMPI-2), mood states were evaluated via both the State-Trait Anxiety Inventory (STAI) and The Center for Epidemiologic Studies Depression Scale (CES-D). Statistical analysis was performed using paired t-tests and Spearman's correlation analyses. The recipient group showed significantly higher scores for Hypochondriasis (t = - 4.49, p = .0001), Depression (t = - 3.36, p = .0015), and Hysteria (t = - 3.30, p = .0018) of MMPI-2 and CES-D (t = - 3.93, p = .0003) than the donor group. The biologically unrelated recipient group reported higher scores of Hypochondriasis (t = - 3.37, p = .003) and Depression (t = - 2.86, p = 0.0098) than the unrelated donor group. Higher scores for Hypochondriasis (t = - 3.00, p = 0.0054) and CES-D (t = - 3.53, p = .0014) were found in the related recipient group. A positive association was found for Hypomania (r = .40, p = .003) of MMPI-2, STAI-S (r = .36, p = .009), and CES-D (r = .36, p = .008) between the recipient and donor groups. Recipients suffered from a higher level of depression and somatic concerns than donors before living kidney transplantation. Psychological problems like depression and anxiety can occur in both living kidney transplantation donors and recipients. This study suggests that clinicians must pay attention to mood states not only in recipients but also in donors because of emotional contagion.


XABOOM: An X-ray Absorption Benchmark of Organic Molecules Based on Carbon, Nitrogen, and Oxygen 1s → π* Transitions.
The performance of several standard and popular approaches for calculating X-ray absorption spectra at the carbon, nitrogen, and oxygen K-edges of 40 primarily organic molecules up to the size of guanine has been evaluated, focusing on the low-energy and intense 1s → π* transitions. Using results obtained with CVS-ADC(2)-x and fc-CVS-EOM-CCSD as benchmark references, we investigate the performance of CC2, ADC(2), ADC(3/2), and commonly adopted density functional theory (DFT)-based approaches. Here, focus is on precision rather than on accuracy of transition energies and intensities-in other words, we target relative energies and intensities and the spread thereof, rather than absolute values. The use of exchange-correlation functionals tailored for time-dependent DFT calculations of core excitations leads to error spreads similar to those seen for more standard functionals, despite yielding superior absolute energies. Long-range corrected functionals are shown to perform particularly well compared to our reference data, showing error spreads in energy and intensity of 0.2-0.3 eV and ∼10%, respectively, as compared to 0.3-0.6 eV and ∼20% for a typical pure hybrid. In comparing intensities, state mixing can complicate matters, and techniques to avoid this issue are discussed. Furthermore, the influence of basis sets in high-level ab initio calculations is investigated, showing that reasonably accurate results are obtained with the use of 6-311++G**. We name this benchmark suite as XABOOM (X-ray absorption benchmark of organic molecules) and provide molecular structures and ground-state self-consistent field energies and spectroscopic data. We believe that it provides a good assessment of electronic structure theory methods for calculating X-ray absorption spectra and will become useful for future developments in this field.


Sarcopenia is associated with incident albuminuria in patients with type 2 diabetes: A retrospective observational study.
Sarcopenia, defined as age-related loss of skeletal muscle mass and function, increases the risk of albuminuria. However, it has still unknown whether sarcopenia could increase the risk for the progression of albuminuria. A total 238 patients with type 2 diabetes (mean age 64 ± 12 years; 39.2% women) were studied in the present retrospective observational study. The prevalence of sarcopenia was 17.6%. During the median follow-up period of 2.6 years, albuminuria was measured 5.8 ± 1.8 times, and progression of albuminuria was observed in 14.9% of patients with normoalbuminuria, as was 11.5% in those with microalbuminuria. Sarcopenia was significantly associated with both progression (hazard ratio 2.61, 95% confidence interval 1.08-6.31, P = 0.034) and regression (hazard ratio 0.23, 95% confidence interval 0.05-0.98, P = 0.048) of albuminuria by multivariate Cox regression analysis. The present data suggest that sarcopenia is an important determinant of both progression and regression of albuminuria in patients with type 2 diabetes.


[Intestinal cryptosporidiosis: a rare diarrheal disease in man].
Report on a 12-year-old boy with diffuse abdominal pains, vomiting and lasting diarrhoea. All the usual bacteriological examinations for Salmonella, Shigella and Campylobacter were negative. Routine search for intestinal parasites by the MIF method was also negative. Cryptosporidium oocysts were, exceptionally, detected in stool smears from the two samples stained by the modified Ziehl-Neelsen method.


Calreticulin-2 is localized in the lumen of the endoplasmic reticulum but is not a Ca2+ -binding protein.
Calreticulin (CRT)-1 is a major Ca(2+)-buffering protein in the lumen of the endoplasmic reticulum. Human and murine CRT-2 was isolated in 2002, but the subcellular localization and function is still unclear. Here, we studied the intracellular localization and function of CRT-2 with hemagglutinin-tagged (HA-) human CRT-2. Western blotting revealed HA-CRT-2 as a single band at 50 kDa. Using immunofluorescence microscopy of cultured fibroblasts and epithelial cells transfected with HA-CRT-2 cDNA, labeling for HA-CRT-2 was seen as a reticular network with a nuclear envelope pattern that colocalized with calnexin and protein disulfide isomerase. Immunoelectron microscopy confirmed that HA-CRT-2 was localized in the lumen of the endoplasmic reticulum. Stains-all staining, a method to detect Ca(2+)-binding proteins, could not stain the immunoprecipitate of HA-CRT-2, although HA-CRT-1 immunoprecipitate was stained blue. These results indicate that the molecular weight of the non-tagged CRT-2 on SDS-PAGE is 49 kDa, and that CRT-2, as well as CRT-1, is localized in the lumen of the endoplasmic reticulum, but that CRT-2 capacity for Ca(2+)-binding may be absent or much lower than that of CRT-1.


mTOR Complex 1 Content and Regulation Is Adapted to Animal Longevity.
Decreased content and activity of the mechanistic target of rapamycin (mTOR) signalling pathway, as well as the mTOR complex 1 (mTORC1) itself, are key traits for animal species and human longevity. Since mTORC1 acts as a master regulator of intracellular metabolism, it is responsible, at least in part, for the longevous phenotype. Conversely, increased content and activity of mTOR signalling and mTORC1 are hallmarks of ageing. Additionally, constitutive and aberrant activity of mTORC1 is also found in age-related diseases such as Alzheimer's disease (AD) and cancer. The downstream processes regulated through this network are diverse, and depend upon nutrient availability. Hence, multiple nutritional strategies capable of regulating mTORC1 activity and, consequently, delaying the ageing process and the development of age-related diseases, are under continuous study. Among these, the restriction of calories is still the most studied and robust intervention capable of downregulating mTOR signalling and feasible for application in the human population.


Systems thinking: A turning point for improving respectful obstetric care in South African health districts.
Poorly functioning health systems and local health systems barriers affect many women giving birth in low- and middle-income countries. The district clinical specialist teams in South Africa are uniquely positioned to provide facilitation and mentoring during interventions for improving the weak primary healthcare system. To ensure success, four key principles should be considered during scale-up of interventions: systems thinking and awareness of contexts and barriers; a focus on sustainability; harnessing factors known to enhance scalability; and respect for human rights and equity. Asking the right questions about the responsibilities of health systems at the micro-, meso- and macro-levels will benefit scale-up processes and sustain innovative pathways to high-quality obstetric care in communities.


[Analysis of the effects of mutant hairless genes in chimeric mice].
The autosomal recessive gene hairless (hr) is responsible for the complete hairlessness in mice homozygous for this gene. Hair shedding that begins at the age of 10 days is caused by an abnormal cycle of hair follicle development disturbed at the catagen stage. This results in enhanced programmed cell death (apoptosis) and ultimately leads to the complete hair follicle destruction and shedding of all hairs by the age of three weeks. To study the phenotypic expression of the hr gene in a chimeric organism, we have obtained 12 chimeric mice hr/hr <--> +/+ by means of aggregation of early embryos hr/hr and +/+. In chimeric mice, the hair shedding has begun two days later than in the hr/hr mice. By day 23 of postnatal development, hairless areas were present on the coat of chimeric mice or the latter were completely hairless depending on the percentage of the hr/hr mutant component. In four chimeras with high content of the mutant component (68-76%), the hair shedding process was similar to that in the hr/hr mice, though it was accomplished two days later. In three chimeras with 48-51% of the mutant component, alternating hairless and hair-covered bands were observed. These data suggest that the hr gene acts in epidermal cells of a hair follicle, because epidermal cell clones in embryonic skin migrate in the lateral-ventral direction coherently and without mixing. However, some chimeras displayed a pattern which was not so clear-cut: the band borders were illegible and hairs partly covered the hairless areas. In some chimeras, the uniform thinning of the coat was observed. Analysis of the effects of the hr mutant gene in chimeric mice differing in the ratio between mutant (hr/hr) and normal (+/+) components in tissues suggests that the hr gene acts in the epidermal cells of the hair follicle. The interactions between cells have an essential effect on the mode and degree of the hr gene expression, which leads to distortion of the "ectodermal" coat pattern in chimeras.


Food frequency questionnaire results correlate with metabolic control in insulin-treated veterans with type 2 diabetes: the Diabetes Outcomes in Veterans Study.
To validate food frequency analysis as a predictor of metabolic status in persons with type 2 diabetes and to identify psychosocial factors affecting dietary adherence. Three hundred forty-seven subjects with stable, insulin-treated type 2 diabetes completed a food frequency questionnaire and six instruments measuring different psychosocial attributes. Eight metabolic parameters were used as principal endpoints. Data from the food frequency questionnaires were used to estimate daily energy intake and determine each subject's adherence to seven dietary standards derived from the 2003 Dietary Recommendations of the American Diabetes Association. We excluded 105 subjects reporting daily energy intake <1,000 kcal because these results were considered unreliable. Subjects were then categorized into three groups depending on their adherence rates to American Diabetes Association dietary standards. Subjects with the lowest dietary adherence had the poorest metabolic control. Adherence to dietary standards was particularly poor among subjects with metabolic syndrome (2.1%) and/or obesity (4.1%). The most important determinants of following recommended dietary practices were positive attitudes, fewer social barriers, and a conviction that diet could control diabetes. For subjects with type 2 diabetes reporting a dietary intake of >1,000 kcal/day, food frequency questionnaire data could identify those with poor metabolic control. Nutrition interventions to improve metabolic control should focus on reducing fat intake and emphasizing the importance of diet. Multidisciplinary efforts should be directed at overcoming social barriers to recommended dietary practices and to treating depression.


Overtraining, excessive exercise, and altered immunity: is this a T helper-1 versus T helper-2 lymphocyte response?
Overtraining syndrome (OTS) occurs where an athlete is training vigorously, yet performance deteriorates. One sign of OTS is suppressed immune function, with an increased incidence of upper respiratory tract infection (URTI). An increased incidence of URTIs is also associated with high volume/intensity training, as well as with excessive exercise (EE), such as a marathon, manifesting between 3-72 hours post-race. Presently, there is no encompassing theory to explain EE and altered immune competence. Recently, it has been conclusively established that T helper lymphocytes (T(H)), a crucial aspect of immune function, represent two distinct functional subsets: T(H)1 and T(H)2 lymphocytes. T(H)1 lymphocytes are associated with cell-mediated immunity (CMI) and the killing of intracellular pathogens, while T(H)2 lymphocytes are associated with humoral immunity and antibody production. When T(H)-precursor cells are activated, the balance is tipped in favour of one or the other. Furthermore, the most appropriate means of determining the T(H)-subset, is by the prevailing cytokine 'pattern'. This paper hypothesises that exercise-related immunosuppression is due to tissue trauma sustained during intense exercise, producing cytokines, which drive the development of a T(H)2 lymphocyte profile. A T(H)2 cell response results in simultaneous suppression of CMI, rendering the athlete susceptible to infection. Additionally, increased levels of circulating stress hormones (cortisol and catecholamines), as well as prostaglandin E(2), support up-regulation of T(H)2 lymphocytes. Marathon-related data are presented to support this hypothesis. It is concluded that an increased incidence of illness associated with OTS and in response to EE is not due to immunosuppression per se, but rather to an altered focus of immune function, with an up-regulation of humoral immunity and suppression of CMI.


Subjective effect of September 11, 2001 among pregnant women: is cumulative history of interpersonal violence important?
Prior history of trauma may sensitize individuals to subsequent trauma, including terrorist attacks. Using a convenience sample of secondary, cross-sectional data, pregnant women were grouped based on lifetime interpersonal violence history. Cumulative risk theory was used to evaluate the association of lifetime interpersonal violence history and subjective impact of the September 11, 2001 (9/11) terrorists attacks. Using hierarchical linear regression, cumulative risk theory was partially supported. Women with a history of only one type of interpfersonal violence reported greater effect of 9/11 than did women without a history, but women with both types of violence did not report a greater effect of 9/11 compared to women endorsing history of one type. These data corroborate the literature in that level of exposure to terrorist-related trauma predicts subjective reaction to the attacks. Future research with a larger sample and standardized instruments is warranted.


CONFLICTS OF INTEREST.
Guest Editors for the "Student Perspectives During COVID-19" section were Roger Gonzales, Jaimee Watts Isley, Wayne R. Lawrence, Jennifer Drey, Brady Rowe, Emily Q. Ritter, AJPH Think Tank; and Paulina Sosa, AJPH Assistant Editor.


Conformational determinants in receptor recognition of peptide hormones: interaction of parathyroid hormone with the glucagon receptor.
Receptor binding assays demonstrate that bovine parathyroid hormone (PTH) and human PTH(1-34) can displace [125I]iodoglucagon from binding to its receptor in rat liver plasma membranes. The displacement of [125I]iodoglucagon requires several thousand-fold more bovine PTH or human PTH(1-34) than glucagon. However, the PTH peptides are more effective than secretin, which up to a concentration of 10(-5) M exhibits no ability to displace [125I]iodoglucagon. The greater potency of PTH compared with secretin occurs despite the fact that secretin shows a great deal of sequence homology with glucagon while PTH shows none. We demonstrate by circular dichroism that in the presence of 3 mM SDS glucagon and hPTH(1-34) have similar secondary structure contents, while secretin is more helical. Our results suggest that receptors can recognize gross conformational features of a peptide hormone in addition to interacting with a specific amino acid sequence. The ability of PTH to interact with glucagon receptors can be modulated by incorporation of charged amphiphiles into the plasma membrane. Negatively charged taurodeoxycholic acid increases the binding of the more cationic PTH while positively charged myristyltrimethylammonium bromide decreases this interaction. These effects demonstrate that receptor specificity can be modulated by its lipid environment and that electrostatic interactions between the hormone and the membrane surface can contribute to receptor binding.


Effects of intensified insulin treatment on retinal vessels in diabetic patients.
Forty-five diabetic patients were randomly assigned to treatment with continuous subcutaneous insulin infusion (CSII), multiple injections (MI), and conventional insulin treatment (CIT). They were prospectively followed up for one year. A computerised scanning microdensitometer was applied on fundus photographs of retinal vessels, and we studied changes in calibres of the blood column (W0) and in width (Wr/W0) and intensity (Ir) of the central 'light reflex'. After six months of improved metabolic control the Ir was reduced in both MI and CSII cases compared with CIT cases (p less than 0.01), indicating haemorrheological changes in the retinas. Within these six months cotton-wool spots appeared in half the patients (n = 15) on CSII and MI, but not in CIT patients. Subjects who developed cotton-wool spots, compared with those who did not, had greater intensities of reflection and larger calibres of vessels at the start of the study (p less than 0.01). On intensifying the treatment they were characterised by a larger fall in hemoglobin A1 (p less than 0.01) and by a larger decrease in Ir on arteries (p less than 0.05) and veins (p less than 0.01). The behaviour of the retinal circulation is different in patients developing transient ischaemic lesions on intensified insulin treatment from its behaviour in those who do not.


Biophysical studies on the stability of DNA intrastrand cross-links of transplatin.
Clinically ineffective transplatin [trans-diamminedichloridoplatinum(II)] is used in the studies of the structure-pharmacological activity relationship of platinum compounds. In addition, a number of transplatin analogs exhibit promising toxic effects in several tumor cell lines including those resistant to conventional antitumor cisplatin. Moreover, transplatin-modified oligonucleotides have been shown to be effective modulators of gene expression. Owing to these facts and because DNA is also considered the major pharmacological target of platinum complexes, interactions between transplatin and DNA are of great interest. We examined, using biophysical and biochemical methods, the stability of 1,3-GNG intrastrand cross-links (CLs) formed by transplatin in short synthetic oligodeoxyribonucleotide duplexes and natural double-helical DNA. We have found that transplatin forms in double-helical DNA 1,3-GNG intrastrand CLs, but their stability depends on the sequence context. In some sequences the 1,3-GNG intrastrand CLs formed by transplatin in double-helical DNA readily rearrange into interstrand CLs. On the other hand, in a number of other sequences these intrastrand CLs are relatively stable. We show that the stability of 1,3-GNG intrastrand CLs of transplatin correlates with the extent of conformational distortion and thermodynamic destabilization induced in double-helical DNA by this adduct.


Dysregulation of the epithelial barrier by environmental and other exogenous factors.
The "epithelial barrier hypothesis" proposes that the exposure to various epithelial barrier-damaging agents linked to industrialization and urbanization underlies the increase in allergic diseases. The epithelial barrier constitutes the first line of physical, chemical, and immunological defense against environmental factors. Recent reports have shown that industrial products disrupt the epithelial barriers. Innate and adaptive immune responses play an important role in epithelial barrier damage. In addition, recent studies suggest that epithelial barrier dysfunction plays an essential role in the pathogenesis of the atopic march by allergen sensitization through the transcutaneous route. It is evident that external factors interact with the immune system, triggering a cascade of complex reactions that damage the epithelial barrier. Epigenetic and microbiome changes modulate the integrity of the epithelial barrier. Robust and simple measurements of the skin barrier dysfunction at the point-of-care are of significant value as a biomarker, as recently reported using electrical impedance spectroscopy to directly measure barrier defects. Understanding epithelial barrier dysfunction and its mechanism is key to developing novel strategies for the prevention and treatment of allergic diseases. The aim of this review is to summarize recent studies on the pathophysiological mechanisms triggered by environmental factors that contribute to the dysregulation of epithelial barrier function.


Food Insecurity Was Associated with Lower Fruits and Vegetables Consumption but Not with Overweight and Obesity in Children from Mexican Fishing Communities.
Food insecurity has been associated with decreased household food supplies, which could affect children's fruit and vegetables intakes and increases their risk of being overweight or obese. The aim of this study was to determine if food insecurity was associated with lower fruit and vegetables consumption and overweight and obesity in children from Mexican fishing communities. A cross-sectional study was 4conducted with 100 elementary-school children aged 6-12 years. A survey that evaluated socioeconomic conditions, food insecurity, and fruits and vegetables consumption was applied. Weight and height were measured. The association between key variables and food insecurity was assessed using linear regression. Sixty-four percent of households experienced food insecurity. Food-insecure children reported lower consumption of fruits and fruits plus vegetables than did their peers with food security (P trend = 0.040 and 0.038, respectively). Food insecurity was also associated with lower chance to meet the recommendation of fruit and vegetable consumption (P trend in logit < 0.001), but was negatively associated with the prevalence of overweight and obesity (P trend in logit < 0.001). Food-insecure children had lower fruits and vegetables consumption but were no more likely to be overweight or obese than their food-secure counterparts.


Contact allergens in ultraviolet-cured acrylic resin systems.
Ultraviolet-cured acrylic resin systems expose the worker to an ultraviolet reactive prepolymer, multifunctional acrylate esters, and a photoinitiator system. Irritation occurs primarily with exposure to the acrylate compounds and increases with an increased number of acrylic double bonds, decreased viscosity, and the presence of hydrophilic groups. It is recommended that sensitive individuals handle these materials with nitrile gloves, but acrylics have been known to penetrate even this form of protection.


Differential rates of local and global homogenization in centromere satellites from Arabidopsis relatives.
Higher eukaryotic centromeres contain thousands of satellite repeats organized into tandem arrays. As species diverge, new satellite variants are homogenized within and between chromosomes, yet the processes by which particular sequences are dispersed are poorly understood. Here, we isolated and analyzed centromere satellites in plants separated from Arabidopsis thaliana by 5-20 million years, uncovering more rapid satellite divergence compared to primate alpha-satellite repeats. We also found that satellites derived from the same genomic locus were more similar to each other than satellites derived from disparate genomic regions, indicating that new sequence alterations were homogenized more efficiently at a local, rather than global, level. Nonetheless, the presence of higher-order satellite arrays, similar to those identified in human centromeres, indicated limits to local homogenization and suggested that sequence polymorphisms may play important functional roles. In two species, we defined more extensive polymorphisms, identifying physically separated and highly distinct satellite types. Taken together, these data show that there is a balance between plant satellite homogenization and the persistence of satellite variants. This balance could ultimately generate sufficient sequence divergence to cause mating incompatibilities between plant species, while maintaining adequate conservation within a species for centromere activity.


Invited commentary: interpreting associations between exposure biomarkers and pregnancy outcome.
Levels of exposure biomarkers vary among individuals because of differences in both environmental exposure and metabolism. However, the ultimate interest is in providing information about the impact of modifying environmental exposures through regulation or behavior change. Using these levels in studies of pregnancy outcomes, as nicely illustrated by the study of Kadhel et al. in this issue of the Journal (Am J Epidemiol. 2014;179(5):536-544), has the usual strength of being integrative across multiple pathways but may reflect reverse causality, in which the underlying disease alters biomarker levels or shared physiological determinants of the biomarker level and the health outcome. Specifically, biomarkers may vary because of spatial differences in exposure, behavioral differences affecting exposure, and metabolic differences across members of the study population. Proper interpretation of such studies calls for a clearer understanding the sources of variation in exposure to more fully consider confounding and reverse causality due to metabolic differences within the study population.


State school nutrition and physical activity policy environments and youth obesity.
With the epidemic of childhood obesity, there is national interest in state-level school policies related to nutrition and physical activity, policies adopted by states, and relationships to youth obesity. This study develops a comprehensive state-level approach to characterize the overall obesity prevention policy environment for schools and links the policy environments to youth obesity for each state. Using 2006 School Health Policies and Programs Study (SHPPS) state data, qualitative and quantitative methods were used (2008-2009) to construct domains of state-level school obesity prevention policies and practices, establish the validity and reliability of the domain scales, and examine their associations with state-level obesity prevalence among youth aged 10-17 years from the 2003 National Survey of Children's Health. Nearly 250 state-level obesity prevention-policy questions were identified from the SHPPS. Three broad policy topic areas containing 100 food service and nutrition (FSN) questionnaire items; 146 physical activity and education (PAE) items; and two weight assessment (WA) items were selected. Principal components analysis and content validity assessment were used to further categorize the items into six FSN, ten PAE, and one WA domain. Using a proportional scaled score to summarize the number of policies adopted by states, it was found that on average states adopted about half of the FSN (49%), 38% of the PAE, and 17% of the WA policies examined. After adjusting for state-level measures of ethnicity and income, the average proportion of FSN policies adopted by states was correlated with the prevalence of youth obesity at r =0.35 (p=0.01). However, no correlation was found between either PAE or WA policies and youth obesity (PAE policies at r =0.02 [p=0.53] and WA policies at r =0.16 [p=0.40]). States appear to be doing a better job adopting FSN policies than PA or WA policies, and adoption of policies is correlated with youth obesity. Continued monitoring of these policies seems to be warranted.


Binding mode of an α-amino acid-linked quinoxaline-2,3-dione analogue at glutamate receptor subtype GluK1.
Two α-amino acid-functionalized quinoxalines, 1a (CNG-10301) and 1b (CNG-10300), of a quinoxaline moiety coupled to an amino acid moiety were designed, synthesized, and characterized pharmacologically. While 1a displayed low affinity at native AMPA, KA, and NMDA receptors, and at homomeric GluK1,3 receptors, the affinity for GluK2 was in the midmicromolar range (Ki = 136 μM), 1b displayed low to midmicromolar range binding affinity at all the iGluRs (Ki = 9-126 μM). In functional experiments (outside-out patches excised from transfected HEK293T cells), 100 μM 1a partially blocked GluK1 (33% peak response), while GluK2 was unaffected (96% peak response). Furthermore, 1a was shown not to be an agonist at GluK1 and GluK2 at 100 μM. On the other hand, 100 μM 1b fully antagonized GluK1 (8% peak response) but only partially blocked GluK2 (33% peak response). An X-ray structure at 2.3 Å resolution of 1b in the GluK1-LBD (ligand-binding domain) disclosed an unexpected binding mode compared to the predictions made during the design phase; the quinoxaline moiety remains to act as an amino acid bioisostere, but the amino acid moiety is oriented into a new area within the GluK1 receptor. The structure of the GluK1-LBD with 1b showed a large variation in domain openings of the three molecules from 25° to 49°, demonstrating that the GluK1-LBD is capable of undergoing major domain movements.


Eelgrass (Zostera marina L.) populations are threatened by high sea-surface temperatures and impaired waters on Nantucket Island, USA.
Eelgrass (Zostera marina L.) is a key foundation species that provides multiple ecosystem services to shallow coastal and estuarine systems in the Northern Hemisphere. It is estimated that, over the last century, up to 50 % of all Z. marina habitat has been lost along the east coast of the USA due to factors including light reduction, eutrophication, and physical disturbance. Warming sea surface temperatures are also believed to be exacerbating losses and the future of this ecosystem is unclear. Here, we assess Z. marina meadows on Nantucket, an island system located 50 km off-shore of Massachusetts, by using common indicators of seagrass plant health and environmental quality. Our results show that Z. marina meadows on Nantucket Island are thermally stressed and light-limited during parts of their peak growing season. This suggests that sea-surface temperatures are a pivotal factor, along with cultural eutrophication, in observed large-scale losses of Z. marina and that further degradation could be expected in the future as the climate continues to warm. Methods from this study may be used by managers as a guide to assess seagrass ecosystem status in degrading systems.


Tuning of a neuro-fuzzy controller by genetic algorithm.
Due to their powerful optimization property, genetic algorithms (GAs) are currently being investigated for the development of adaptive or self-tuning fuzzy logic control systems. This paper presents a neuro-fuzzy logic controller (NFLC) where all of its parameters can be tuned simultaneously by GA. The structure of the controller is based on the radial basis function neural network (RBF) with Gaussian membership functions. The NFLC tuned by GA can somewhat eliminate laborious design steps such as manual tuning of the membership functions and selection of the fuzzy rules. The GA implementation incorporates dynamic crossover and mutation probabilistic rates for faster convergence. A flexible position coding strategy of the NFLC parameters is also implemented to obtain near optimal solutions. The performance of the proposed controller is compared with a conventional fuzzy controller and a PID controller tuned by GA. Simulation results show that the proposed controller offers encouraging advantages and has better performance.


Updates on the treatment of essential hypertension: a summary of AHRQ's comparative effectiveness review of angiotensin-converting enzyme inhibitors, angiotensin II receptor blockers, and direct renin inhibitors.
In 2007, the Agency for Healthcare Research and Quality (AHRQ) published a comparative effectiveness review (CER) on the benefits and risks of angiotensin-converting enzyme inhibitors (ACEIs) and angiotensin II receptor blockers (ARBs) for treating essential hypertension in adults. The main findings indicated that the 2 classes of antihypertensive medications caused similar reductions in blood pressure, although higher rates of adverse events, especially cough, were reported by patients treated with ACEIs. In addition, the 2007 review indicated no treatment related differences in lipid levels, glycemic control, or progression of kidney disease among the agents. Since 2007, 39 relevant studies have been published that compare outcomes for adults treated with ACEIs versus ARBs or a drug in one of these 2 classes versus a direct renin inhibitor (DRI). To systematically analyze findings from the new research, AHRQ commissioned and, in June 2011, published an updated comparative effectiveness review on the benefits and risks of agents that target the renin-angiotensin- aldosterone system (RAAS), specifically ACEIs, ARBs, and DRIs. To (a) familiarize health care professionals with the methods and findings from AHRQ's 2011 comparative effectiveness review on ACEIs, ARBs, and DRIs for adults with essential hypertension; (b) provide commentary and encourage consideration of the clinical and managed care applications of the review findings; and (c) identify limitations to the existing research on the benefits and risks of ACEIs, ARBs, and DRIs. Consistent with the findings from AHRQ's 2007 report, the 2011 update indicated no overall differences in blood pressure control, mortality rates, and major cardiovascular events in patients treated with ACEIs versus ARBs. With a low strength of evidence, 2 studies reported a small significantly greater blood pressure reduction for patients treated with the DRI aliskiren versus the ACEI ramipril. Studies evaluating the DRI aliskiren versus ACEIs and ARBs on mortality and morbidity outcomes were relatively short, and few deaths or cardiovascular events occurred, resulting in insufficient evidence to discern differences. A random-effects meta-analysis of 23 RCTs comparing ACEIs and ARBs found no significant difference in the proportion of patients who achieved successful blood pressure control on a single antihypertensive agent. Compared with ARBs and the DRI aliskiren, ACEIs were consistently associated with higher rates of cough. Withdrawals due to adverse events were modestly more frequent for patients receiving ACEI rather than ARBs or DRIs; this is consistent with the differential rates of cough. There was no evidence of differential effects of ACEIs, ARBs, or DRIs on the outcomes of lipids, renal outcomes, carbohydrate metabolism or diabetes, or left ventricular mass; however, there was not a high strength of evidence for any of these outcomes. Regarding the question of whether ACEIs, ARBs, or DRIs are associated with better outcomes in specific patient subgroups, the evidence was insufficient to reach firm conclusions.


Broad Virus Detection and Variant Discovery in Fecal Samples of Hematopoietic Transplant Recipients Using Targeted Sequence Capture Metagenomics.
Pediatric allogeneic hematopoietic stem cell transplantation (HSCT) patients often suffer from gastro-intestinal (GI) disease caused by viruses, Graft-versus-Host Disease (GVHD) or a combination of the two. Currently, the GI eukaryotic virome of HSCT recipients remains relatively understudied, which complicates the understanding of its role in GVHD pathogenicity. As decisions regarding immunosuppressive therapy in the treatment of virus infection or GVHD, respectively, can be completely contradicting, it is crucial to better understand the prevalence and relevance of viruses in the GI tract in the HSCT setting. A real time PCR panel for a set of specific viruses widely used to diagnose the most common causes of GI viral gastroenteritis is possibly insufficient to grasp the full extent of viruses present. Therefore, we applied the targeted sequence capture method ViroCap to residual fecal samples of 11 pediatric allogeneic HSCT recipients with GI symptoms and a suspicion of GVHD, to enrich for nucleic acids of viruses that are known to infect vertebrate hosts. After enrichment, NGS was applied to broadly detect viral sequences. Using ViroCap, we were able to detect viruses such as norovirus and adenovirus (ADV), that had been previously detected using clinical diagnostic PCR on the same sample. In addition, multiple, some of which clinically relevant viruses were detected, including ADV, human rhinovirus (HRV) and BK polyomavirus (BKV). Interestingly, in samples in which specific PCR testing for regular viral GI pathogens did not result in a diagnosis, the ViroCap pipeline led to the detection of viral sequences of human herpesvirus (HHV)-7, BKV, HRV, KI polyomavirus and astrovirus. The latter was an only recently described variant and showed extensive sequence mismatches with the applied real time PCR primers and would therefore not have been detected if tested. Our results indicate that target enrichment of viral nucleic acids through ViroCap leads to sensitive and broad possibly clinically relevant virus detection, including the detection of newer variants in clinical HSCT recipient samples. As such, ViroCap could be a useful detection tool clinically, but also in studying the associations between viral presence and GVHD.


Mortality in Micronesian Nauruans and Melanesian and Indian Fijians is not associated with obesity.
The association of obesity with mortality was investigated in population-based samples of Micronesian Nauruans (n = 1,400), Melanesian Fijians (n = 1,279), and Indian Fijians (n = 1,182), over 10 years from 1982 in Nauru, and 11 years from 1980 in Fiji. At the end of follow-up, vital status was known for all Nauruans and all but 3.5% of Fijians. Mortality rates were higher in Nauru than Fiji, and in Melanesians than Indians. The mean body mass index of decedents was similar to or less than (Nauruan men, p < 0.001) that of survivors in each sex-ethnic group. Crude mortality rates showed an inverse relation with body mass index in Nauruan men, with inconsistent relations in other sex-ethnic groups. After stratification by diabetes status, there was no relation between mortality and obesity in nondiabetic subjects, but an inverse relation was observed among diabetic subjects in each population. These findings persisted even after the exclusion of subjects who died within the first 2 years of follow-up. After controlling for age, smoking, and diabetes status in Cox proportional hazard models, body mass index (as a continuous variable) was not related to mortality in any sex/ethnic group and tended to be negatively associated with mortality risk. Interactions of body mass index with age, smoking, and diabetes status were not significant. Mortality risk was significantly increased in older subjects and in diabetic subjects, and cigarette smoking also increased risk in some groups. Stratification of analyses according to cigarette smoking did not alter the nature of the results. The association of mortality and body mass index categorized by quartiles was also investigated. After adjusting for age alone, or age, smoking, and diabetes status, the lower quartiles of body mass index were consistently associated with the highest relative risk for mortality. Quadratic terms for body mass index did not improve Cox models in subjects with normal glucose tolerance. Relations with cardiovascular disease mortality were also assessed and results were inconsistent, although positive trends were observed in Nauruan women (p = 0.02) and Melanesian men (p = 0.06). Overall, there was little evidence to suggest that obesity was a risk factor for total or cardiovascular mortality in these populations. However, obesity is clearly associated with a high risk of diabetes and other morbid conditions and at least on this basis it would seem desirable to prevent obesity in these and other Pacific populations.


Making bigger brains-the evolution of neural-progenitor-cell division.
Relative brain size differs markedly between species. This variation might ultimately result from differences in the cell biology of neural progenitors, which might underlie their different proliferative potential. On the basis of the cell-biological properties of neural progenitors of animals of varying brain size and complexity (namely, Drosophila melanogaster, rodents and primates), we hypothesize that the evolution of four related cell-biological features has contributed to increases in neuron number. Three of these features-the pseudostratification of the progenitor layer, the loss of (Inscuteable-mediated) mitotic-spindle rotation and the evolution of proteins (such as Aspm) that maintain the precision of symmetric progenitor division-affect the mode of cell division in the apically dividing progenitors of the ventricular zone. The fourth feature, however, concerns the evolution of the basally dividing progenitors of the subventricular zone. In rodents, these basal (or intermediate) progenitors lack cell polarity, whereas in primates a subpopulation of radial, presumably polarized, progenitors has evolved (outer-subventricular-zone progenitors). These cells undergo basal mitoses and are thought to retain epithelial characteristics. We propose the epithelial-progenitor hypothesis, which argues that evolutionary changes that promote the maintenance of epithelial features in neural progenitors, including outer-subventricular-zone progenitors, have been instrumental in the expansion of the cerebral cortex in primates.


MTHFR C677T genotype and cardiovascular risk in a general population without mandatory folic acid fortification.
Meta-analyses have suggested an effect of MTHFR C677T genotype (rs1801133), a proxy for blood total homocysteine, on cardiovascular disease (CVD) in populations with low population dietary folate. The aim was to examine the association and effect modification by serum folate and vitamin B12 levels between MTHFR and CVD-related outcomes in a general population with no mandatory folic acid fortification policy. The study population included 13,748 adults retrieved from pooling of four population-based studies conducted in Denmark. MTHFR genotype, serum folate (measured in approximately 9,356 individuals), and serum vitamin B12 (9,215 individuals), hypertension, and dyslipidemia were measured at baseline, and participants were followed for a mean of 10.5-11.7 years in central registries for diagnoses of stroke (623 incidents), ischaemic heart disease (IHD) (835 incidents), and all-cause mortality (1,272 incidents). The MTHFR genotype (TT vs. CC/CT) was not associated with hypertension [OR (95% CI) 1.09 (0.95-1.25)], dyslipidemia [OR (95% CI) 0.97 (0.84-1.11)], stroke [HR (95% CI) 0.92 (0.69-1.23)], and all-cause mortality [HR (95% CI) 0.94 (0.77-1.14)], either overall, or in participants with low serum folate or B12 status (P values for interactions 0.15-0.94). Individuals with the MTHFR TT genotype had a higher risk of IHD (HR (95% CI) 1.38 (1.11-1.71)), but this association was not modified by folate status (P value for interaction 0.45). Our results do not support a causal relationship between homocysteine and CVD. However, we cannot exclude a direct causal effect of MTHFR C677T genotype on IHD.


[The hospital nursing discharge form. Establishing and validation of a standard formate].
The purpose of this article is to describe the design, application and validation process for a new hospital nursing medical discharge form. This process has three phases: FIRST PHASE: Redesigning the Document. In 1996, as a response to growing demands as well as the need to adapt to the requirements of data processing, all nursing documents in all hospital nursing files were updated following the recommendations of the Technical Commission. SECOND PHASE: Initial Application and Validation. The medical discharge form prepared in phase one was applied during the first trimester of 1997. The parties involved agreed to test this form until the end of the year in order to come to a consensus regarding its structure and content while at the same time determining its degree of comprehension and usefulness. THIRD PHASE: Editing and Final Format. The results of these analyses, together with revisions of criteria included in the NMDS and from the Conference on Hospital Discharge Abstract System, made it possible to draw up the final draft of this form which include nursing discharge criteria in the evaluation section. A pilot test was carried out in five hospital units to determine the validity of this form according to the same criteria of comprehension and usefulness. The results indicate a recommendation to eliminate the sections of medical diagnosis and medication upon discharge while to maintain the identification of the patient, the summary of his/her stay in the center, the description of the patient's case upon discharge and the treatment plan.


Effect of hypertonic saline solution on left ventricular afterload in normovolumic dogs.
The effects of hypertonic saline solution (HSS) and hyperosmotic dextrose (HD; 2,400 mosm/L, 4 ml/kg of body weight) on left ventricular afterload were determined in normovolumic, chloralose-anesthetized, autonomically blocked dogs (n = 8). Solutions were infused IV over 3 minutes. Left ventricular afterload was assessed by use of a dual-tipped micromanometer catheter with an electromagnetic fluid-velocity sensor located in the ascending aorta, and the impedance spectrum was calculated after Fourier analysis of signal-averaged aortic pressure and flow signals. Hypertonic saline solution and HD decreased peripheral resistance, reflection coefficient at zero frequency, and frequency of the first zero crossing of the phase angle for 3 to 5 minutes after either fluid was administered. Characteristic impedance was not altered by HSS or HD. These impedance spectrum changes indicate transient vasodilatation and afterload reduction. We conclude that the vascular effect of an ionic hyperosmotic solution (HSS) is similar to that of a nonionic hyperosmotic solution (HD), and that HSS and HD transiently decrease afterload in normovolumic dogs. The duration of the afterload reduction after HSS administration appeared to be too short to be of great clinical benefit.


Community Intervention Trial for Smoking Cessation (COMMIT): summary of design and intervention. COMMIT Research Group.
The Community Intervention Trial for Smoking Cessation (COMMIT) is a multicenter project created to test a comprehensive smoking control intervention. COMMIT was designed to effectively reach cigarette smokers, especially heavy smokers, and to aid them in achieving and maintaining long-term cessation of cigarette smoking. COMMIT was also designed to work through communities, using existing media channels, major organizations, and social institutions capable of influencing smoking behavior in large groups of people. This ongoing trial was begun in 1989 in 11 matched pairs of communities. Each pair consists of one community randomized to intervention conditions and one community randomized to comparison conditions. A comprehensive set of mandated activities is organized around four task forces encompassing health care providers, work sites and organizations, cessation resources and services, and public education. Intervention activities are adapted and implemented in each community through a local community board that provides oversight and management of COMMIT activities. Cessation rates will be assessed in randomly selected cohorts of heavy cigarette smokers, identified at the baseline survey and tracked for 5 years, as estimators of the overall quit rates in both intervention and comparison communities. Various data collection methods will be used to provide intermediate outcome measurement and to evaluate implementation. These methods include cross-sectional community surveys and surveys of health providers, work sites, religious organizations, cessation resource providers, and other systems used to track smoking control activities. The trial will have an impact on more than 200 000 adult smokers.


Parotid abscess: a retrospective study of 14 cases at a regional hospital in Taiwan.
Acute suppurative sialadenitis commonly affects the parotid gland. However, acute suppurative parotitis with abscess formation is less common and possibly complicated by deep neck space infection and sepsis. Our aim was to analyze the clinical features, radiological findings, treatment modalities, and microbiology of parotid abscesses treated at a regional hospital in Taiwan over a 15-year period. Records from patients diagnosed with acute suppurative parotitis or parotid abscesses between January 1998 and December 2012 were retrospectively reviewed. Parotid abscesses were confirmed by computed tomography (CT) examinations. Fourteen patients (9 males; mean age+standard deviation, 49.6 +/- 14.4 [range, 22-75] years) with parotid abscesses were included. Painful swelling at the angle of the jaw was the most common symptom. All patients sustained unilateral parotid abscesses, with left-sided lesions in 10 patients, and right-sided ones in 4 patients. Radiographically, 13 of 14 abscess lesions were located in the superficial lobe, and 1 was in the deep lobe of the parotid gland. Besides sufficient fluid hydration, maintenance of good oral hygiene, and administration of parenteral antibiotics, all patients were treated with surgical incision and drainage. Klebsiella pneumoniae was the organism most commonly isolated from abscess cultures. Parotid abscesses were found in about one fifth of patients with acute infectious parotid disease. Parenteral antibiotics plus surgical incision and drainage was the treatment of choice.


Pediatric Neuro-oncology.
This article reviews the most common pediatric brain tumors, neurocutaneous syndromes, treatment-related neurotoxicities, and the long-term outcomes of survivors. In the era of molecular diagnostics, the classification, management, and prognostication of pediatric brain tumors and neurocutaneous syndromes has been refined, resulting in advancements in patient management. Molecular diagnostics have been incorporated into the most recent World Health Organization 2021 classification. This knowledge has allowed for novel therapeutic approaches targeting the biology of these tumors with the intent to improve overall survival, decrease treatment-related morbidity, and improve quality of life. Advances in management have led to better survival, but mortality remains high and significant morbidity persists. Current clinical trials focus on tumor biology targeted therapy, deescalation of therapy, and multimodal intensified approaches with targeted therapy in more high-risk tumors. Molecular diagnostics for pediatric brain tumors and neurocutaneous syndromes have led to novel therapeutic approaches targeting the biology of these tumors with the goals of improving overall survival and decreasing treatment-related morbidity. Further understanding will lead to continued refinement and improvement of tumor classification, management, and prognostication.


Sleep Disturbances and Nightmares in a Patient Treated with Prazosin.
Prazosin is increasingly being used off-label to treat nightmares in patients with posttraumatic stress disorder. The literature about the psychiatric adverse effects of prazosin is very limited. We present a case in which low-dose prazosin was associated with nightmares and sleep disturbances in an elderly patient without previously diagnosed mental illness or coexisting environmental risk factors for nightmares. Insomnia and hallucinations are listed as some of the rare side effects of prazosin by the manufacturer. Prazosin could be associated with rare psychiatric adverse effects and sleep disturbances. Particular attention is required in identifying these adverse effects, which can be difficult to distinguish from other drug-related side effects in the elderly particularly because they are often using multiple medications.


Lymphatic absorption and tissue disposition of liposome-entrapped [14C]adriamycin following intraperitoneal administration to rats.
The lymphatic absorption and tissue distribution of free [14C]Adriamycin, "empty" [3H]liposomes, free [14C]Adriamycin plus empty [3H]liposomes, and [14C]Adriamycin entrapped into [3H]liposomes have been examined at intervals after i.p. injection into rats. Following treatment with empty [3H]liposomes, almost 30% of the liposomal lipid marker was recovered in 24-hr thoracic duct lymph, but when [14C]Adriamycin was added to or encapsulated in liposomes, this value was reduced to 10%. Conversely, only 1% of free [14C]Adriamycin was recovered in 24-hr lymph, but liposomal encapsulation produced a six-fold increase in this value. Studies on the tissue distribution of the liposomal lipid marker after dosing with empty liposomes revealed uptake by diaphragm, liver and spleen, but the highest tissue concentrations were noted in lymph nodes. Liposomal encapsulation of Adriamycin altered its tissue disposition, chiefly by increasing the concentration of drug equivalents in diaphragm, liver and spleen. Although free Adriamycin was accumulated by lymph nodes to some extent, this lymph node accumulation was markedly enhanced by liposomal encapsulation and was present only in those nodes through which lymph draining the peritoneal cavity passes. This finding, together with the observation that diaphragm and thoracic duct lymph contain relatively high levels of liposomal lipid and Adriamycin equivalents, indicates that liposomes are selectively absorbed from the peritoneal cavity by lymphatics and are retained by certain lymph nodes. The results of this study suggest that i.p. administration of liposome-encapsulated drugs may provide a means of selectively concentrating anti-tumor agents in lymphatic channels and lymph nodes.


Whole-genome analysis of two-component signal transduction genes in fungal pathogens.
Two-component phosphorelay systems are minimally comprised of a histidine kinase (HK) component, which autophosphorylates in response to an environmental stimulus, and a response regulator (RR) component, which transmits the signal, resulting in an output such as activation of transcription, or of a mitogen-activated protein kinase cascade. The genomes of the yeasts Saccharomyces cerevisiae, Schizosaccharomyces pombe, and Candida albicans encode one, three, and three HKs, respectively. In contrast, the genome sequences of the filamentous ascomycetes Neurospora crassa, Cochliobolus heterostrophus (Bipolaris maydis), Gibberella moniliformis (Fusarium verticillioides), and Botryotinia fuckeliana (Botrytis cinerea) encode an extensive family of two-component signaling proteins. The putative HKs fall into 11 classes. Most of these classes are represented in each filamentous ascomycete species examined. A few of these classes are significantly more prevalent in the fungal pathogens than in the saprobe N. crassa, suggesting that these groups contain paralogs required for virulence. Despite the larger numbers of HKs in filamentous ascomycetes than in yeasts, all of the ascomycetes contain virtually the same downstream histidine phosphotransfer proteins and RR proteins, suggesting extensive cross talk or redundancy among HKs.


Prohibitin expression is associated with high grade breast cancer but is not a driver of amplification at 17q21.33.
In a study of ductal carcinoma in situ of the breast, we identified five genes at chromosome 17q21.33 that were over-expressed in high grade cases, and showed a correlation between expression and gene copy number. The aim of this study was to investigate potential drivers of genomic amplification at 17q21.33. Analysis of high resolution comparative genomic hybridisation and published data specified a minimum region of amplification at 17q21.33. Prohibitin (PHB) expression was examined by immunohistochemistry in 285 invasive breast cancers. Gene copy number was examined by fluorescence in situ hybridisation. The minimum region of amplification at 17q21.33 included ten genes with PHB selected as a candidate driver. Increased PHB expression was associated with higher grade breast cancer and poorer survival. Amplification of PHB was detected in 13 of 235 cases (5.5%) but was not associated with PHB expression. PHB amplification was most common in the ERBB2+ breast cancer subtype, although high expression was most prevalent in basal-like and luminal B cancers. Amplification at 17q21.33 is a recurrent feature of breast cancer that forms part of a 'firestorm' pattern of genomic aberration. PHB is not a driver of amplification, however PHB may contribute to high grade breast cancer.


Pareidolias in REM Sleep Behavior Disorder: A Possible Predictive Marker of Lewy Body Diseases?
To investigate conditions and clinical significance of pareidolias in patients with idiopathic rapid eyemovent (REM) sleep behavior disorder (iRBD). This cross-sectional study examined 202 patients with iRBD (66.8 ± 8.0 yr, 58 female) and 46 healthy control subjects (64.7 ± 5.8 years, 14 females). They underwent the Pareidolia test, a newly developed instrument for evoking pareidolias, video polysomnography, olfactory tests, and Addenbrooke's cognitive examination-revised. Results show that 53.5% of iRBD patients exhibited one or more pareidolic responses: The rate was higher than control subjects showed (21.7%). The pictures evoking pareidolic responses were more numerous for iRBD patients than for control subjects (1.2 ± 1.8 vs. 0.4 ± 0.8, p < .001). Subgroup analyses revealed that iRBD patients with pareidolic responses had higher amounts of REM sleep without atonia (RWA), with lower sleep efficiency, lower cognitive function, and older age than subjects without pareidolic responses. Results of multivariate analyses show the number of pareidolic responses as a factor associated with decreased cognitive function in iRBD patients with better predictive accuracy. Morbidity length and severity of iRBD, olfactory function, and the amount of RWA were not factors associated with better predictive accuracy. Half or more of the iRBD patients showed pareidolic responses. The responses were proven to be associated more intimately with their cognitive decline than clinical or physiological variables related to RBD. Pareidolias in iRBD are useful as a predictive marker of future development of Lewy body diseases.


Prevention of urinary tract infection in rats with an indigenous Lactobacillus casei strain.
Our previous studies have shown that indigenous bacteria are able to block the in vitro attachment of uropathogenic bacteria to human uroepithelial cells. In the present study, we applied the concept of competitive exclusion to an animal model. A chronic urinary tract infection was established in female rats with bacteria incorporated into agar beads injected periurethrally into the urinary bladder via a no. 3 French ureteral catheter. Five strains of uropathogenic organisms were used in the first set of experiments, and their colonization of the bladder and kidneys of the animals was confirmed up to 2 months after injection. The uropathogens stimulated an immune response, detected by serum antibodies against the uropathogens, and an inflammatory response noted in sections of the kidneys stained with hematoxylin and eosin. Using this animal model, we established the persistent adherence of bacteria in the urinary tract without the need for creation of obstruction or implantation of a foreign body. In a second set of experiments, an isolate of Lactobacillus casei GR1 taken from the urethra of a healthy woman was incorporated into agar beads, instilled within the rat bladders on day 1, and then swabbed twice weekly for 21 days onto the introitus before challenge with uropathogens instilled into the urinary bladder. In 21 of 25 animals, no uropathogenic bacteria were recovered from the bladder and kidney tissues up to 60 days after challenge, and no immune response was detected. Our results show that L. casei prevented onset of urinary tract infection in 84% of the animals tested. The lactobacilli appeared to exclude the uropathogens from colonizing the urinary tract, within the first 48 hours after challenge, and the net effect was a complete eradication of bacteria from the uroepithelium. It is hoped that the demonstration of a protective role for indigenous bacteria in preventing urinary tract infection in an animal model will lead to the application of this technology to prevent recurrent urinary tract infection in female patients.


Side-dependent effects of internal versus external Na and K on ouabain binding to reconstituted human red blood cell ghosts.
The side-dependent effects of internal and external Na and K on the ouabain binding rate, as promoted by inside MgATP, has been evaluated utilizing reconstituted human red blood cell ghosts. Such ghost systems provide the situation where [Na]i, [K]i, [Na]o, and [K]o can each be varied under conditions in which the others are either absent or fixed at constant concentrations. It was found that, in the presence of Ko, increasing either [Na]i or [K]i resulted in decreasing the rate at which ouabain was bound. Changes in [Na]i or [K]i in the absence of Ko were without effect on the ouabain binding rate. Thus, the ouabain binding rate was found to vary inversely with the rate of Na:K and K:K exchange but was independent of the rate of Na:Na exchange. The effect of Ko in antagonizing ouabain binding, as well as the influence of Nao on this interaction, were found to require the presence of either Nai or Ki. The results are interpreted in terms of a model relating the availability of the ouabain binding site to different conformational states of the pump complex. Differences were observed in the ouabain binding properties of red cell ghosts compared to microsomal preparations but it is not known whether the basis for the differences resides in the different preparations studied or in the lack of control of sidedness in the microsomal systems.


Prospective separation of normal and leukemic stem cells based on differential expression of TIM3, a human acute myeloid leukemia stem cell marker.
Hematopoietic tissues in acute myeloid leukemia (AML) patients contain both leukemia stem cells (LSC) and residual normal hematopoietic stem cells (HSC). The ability to prospectively separate residual HSC from LSC would enable important scientific and clinical investigation including the possibility of purged autologous hematopoietic cell transplants. We report here the identification of TIM3 as an AML stem cell surface marker more highly expressed on multiple specimens of AML LSC than on normal bone marrow HSC. TIM3 expression was detected in all cytogenetic subgroups of AML, but was significantly higher in AML-associated with core binding factor translocations or mutations in CEBPA. By assessing engraftment in NOD/SCID/IL2Rγ-null mice, we determined that HSC function resides predominantly in the TIM3-negative fraction of normal bone marrow, whereas LSC function from multiple AML specimens resides predominantly in the TIM3-positive compartment. Significantly, differential TIM3 expression enabled the prospective separation of HSC from LSC in the majority of AML specimens with detectable residual HSC function.